Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics cDNA Clone
Tissue cDNA, First Strand, Human Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics
Purity
Molecular Biology Grade
Synonyms
Tissue cDNA; First Strand Diseased; Alzheimer's Disease; Brain; Hippocampus; BioGenomics; First Strand; Human Diseased; BioGenomics cdna clone
Purity/Purification
Molecular Biology Grade
Form/Format
Supplied as a liquid in 1 X RT Buffer: 50mM Tris-CI, QH 8.3, 75mM KCl, 3mM MgCl2, lOmM DTT.
Concentration
2.5ng/ul (varies by lot)
Application Notes
Immediate PCR Amplification of known genes, Verification of genetic mutation, Comparison of a specific gene between different tissues, Analysis of mRNA alternative splicing, Gene cloning and target sequencing
Quality Control
1. The Integrity of the RNA used for cDNA syntnesls is examined by' visual inspection for the [2resence of intact bands of 18s and 28s ribosomal RNA when electrophoreses on a denaturing agarose gel. The quality and purity of total RNA were tested by spectrophotometer. A260/280 is between 1.8 and 2.0 (detected in 1 OmM Tris-CI, pH 7.5). The ratio of 28S/18S is =>1.
2. The RNA used for cDNA synthesis is treated by' DNase I, and is tested as DNA free RNA by PCR.
3. The synthesized cDNA was 5' selected to ensure its full length. The cDNA was used as template for PCR amplification of b-actin gene and an 838bp b-actin band was visualized on 2% agarose gel. b-Actin control primer is included.
2. The RNA used for cDNA synthesis is treated by' DNase I, and is tested as DNA free RNA by PCR.
3. The synthesized cDNA was 5' selected to ensure its full length. The cDNA was used as template for PCR amplification of b-actin gene and an 838bp b-actin band was visualized on 2% agarose gel. b-Actin control primer is included.
Unit
1 ul cDNA is sufficient for one PCR reaction.
Donor Information
Human, male, 88 years old, disease, brain, hippocampus.
Clinical Identification
Alzheimer's Disease
Control PCR condition
Ready First Strand eDNA: 1 ul
lOX PCR Buffer: 2.5ul
10mM dNTP: O.5ul
Control Primers (SuM): 1 ul
H20, Nuclease-free: 19.8ul
Taq Polymerase (5u/ul): 0.2ul
PCR Thermocycling:
• 94 degree C x 2 minutes 1 cycle.
• 94 degree C x 30 seconds, 55 degree C x 30 seconds, 72 degree C x 30 seconds, 35 cycles.
• 72 degree C x 5 minutes, 1 cycle. Then hold at 4 degree C
Reaction Mixture:
PCR Ready First Strand cDNA: 1 ul
lOx PCR Buffer : 2.5ul
2.5 mM dNTP: 4ul
Control primers (25uM): 1 ul
H20 Nuclease-free: 16.3ul
Taq Polymerase(5u/ul): 0.2ul
The peR thermocycling:
94 degree C x 2 minutes 1 cycle
94 degree C x 30 seconds, 55 degree C x 45 seconds, 72 degree C x 30 seconds, 35 cycles
72 degree C x 5 minutes, 1 cycle.
Then hold at 4 degree C.
lOX PCR Buffer: 2.5ul
10mM dNTP: O.5ul
Control Primers (SuM): 1 ul
H20, Nuclease-free: 19.8ul
Taq Polymerase (5u/ul): 0.2ul
PCR Thermocycling:
• 94 degree C x 2 minutes 1 cycle.
• 94 degree C x 30 seconds, 55 degree C x 30 seconds, 72 degree C x 30 seconds, 35 cycles.
• 72 degree C x 5 minutes, 1 cycle. Then hold at 4 degree C
Reaction Mixture:
PCR Ready First Strand cDNA: 1 ul
lOx PCR Buffer : 2.5ul
2.5 mM dNTP: 4ul
Control primers (25uM): 1 ul
H20 Nuclease-free: 16.3ul
Taq Polymerase(5u/ul): 0.2ul
The peR thermocycling:
94 degree C x 2 minutes 1 cycle
94 degree C x 30 seconds, 55 degree C x 45 seconds, 72 degree C x 30 seconds, 35 cycles
72 degree C x 5 minutes, 1 cycle.
Then hold at 4 degree C.
Preparation and Storage
Store cDNAs at -20 degree C
Related Product Information for Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics cdna clone
Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques. 1 1 ug total RNA was primed by an oligo dT pnmer and reverse transcribed by MMLV reverse transcriptase in 40ul tinal volume. RT Reaction stor2ped by heating at 65 degree C for 10 minutes. The cDNA is in 1 X RT buffer. (1 X RT Buffer: 50mM Tris-CI, pH 8:1. 75mM KCl, 3mM MgCl2, 10mM D I I). The estimated cDNA concentration is about 2.5ng/ul. 1 ul cDNA is sufficient for one PCR reaction.
Product Categories/Family for Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics cdna clone
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Product Notes
The Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics (Catalog #AAA652008) is a cDNA Clone and is intended for research purposes only. The product is available for immediate purchase. Immediate PCR Amplification of known genes, Verification of genetic mutation, Comparison of a specific gene between different tissues, Analysis of mRNA alternative splicing, Gene cloning and target sequencing. Researchers should empirically determine the suitability of the Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Tissue cDNA, First Strand Diseased, Alzheimer's Disease, Brain, Hippocampus, BioGenomics, cDNA Clone" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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